Figure 4: Allogeneic GFP

(A) Immunohistochemistry identified a small number of donor GFPpos pPICs at 14 days post-transplantation, scale bar = 100 μm. GFPpos cells were found as nuclei within the interstitial spaces between the muscle fibers (bottom inset), and a rare GFPpos muscle fiber was also identified (top inset). (B) Quantification of GFPpos cells to total number of nuclei or myofibers in paraffin embedded skeletal muscle cross sections, n = 5. Data are mean ± SD, *p = 0.001 versus CTX-PBS–treated. (C) Quantification of CD8pos cells per total nuclei (%) determined from 5 fields of view per group. Data are mean ± SD, *p = 0.0008 versus uninjured muscle, **p = 0.0008 versus CTX-PBS–treated. (D) Quantification of Granzyme Bpos cells per total nuclei (%) determined from 5 fields of view per group. Data are mean ± SD, *p < 0.0001 versus uninjured muscle, **p < 0.0001 versus CTX-PBS–treated. (E) Quantification of IBA-1pos cells per total nuclei (%) determined from 5 fields of view per group. Data are mean ± SD, *p = 0.0004 versus uninjured muscle. Porcine PICs were co-cultured with either (F) PHA-activated porcine PBMCs or (G) inactivated porcine PBMCs at different ratios and analyzed for apoptosis using Annexin V/7-AAD staining and flow cytometry. Early apoptosis of CD45neg PICs is represented by Annexin V expression, whereas late apoptosis is represented by double-positive Annexin V/7-AAD staining, determined from duplicate wells per group. Data are mean ± SD, *p < 0.0001 versus pPICs only. PBMC = peripheral blood mononuclear cell; other abbreviations as in Figure 1.